PROJECT SUMMARY - VDDRC FLOW CYTOMETRY CORE The Flow Cytometry and Cell Sorting Core is an established core for the VDDRC. It was developed in direct response to member requests and approved by the Executive Committee and the Scientific Advisory Board with substantial institutional support provided by the University. This core provides access to state-of-the-art equipment for analytical cytometric analysis, cell counts and viability, and cell sorting, and it provides expert training and consulting on the use of these methodologies. Forty-five (45) VDDRC members used this core over the last funding period. The core facility maintains analytical flow cytometers, three cell sorters, an imaging cytometer, a mass cytometer, a mobile cytometer than can be positioned in any lab, and a high-speed plate sampler. The core provides both Mac and PC workstations, an extensive toolbox of software for flow cytometry, and digital backup of data on a secure remote server. Sample biosafety is carefully considered and policies and practices are in place to safely process samples and tissues up to the BSL-3 level. The staff provides both group and one-on-one instruction to facilitate development of acquisition and analysis skills for all users. Trained users can use the analytical cytometers directly on a 24 hour/7 days a week/365 days per year basis; sorting is an assisted-only service during normal operational hours. The core also has worked with VDDRC investigators to develop novel sorting techniques for elements of subcellular compartments such as vesicles. This technically forward work requires custom modification of the sorting cytometer in collaboration with the manufacturer, and involves a large number of technical components from gradient preparations, staining, lasers and fluidics, and downstream collection and proteomics analysis. The core works with VDDRC investigators in cutting edge fields such as Mass Cytometry to identify and analyze rare cells with high-content measurements. The core maintains an active development program to keep the instrumentation and systems current, functional, accessible, and easy to use. In fact, a number of our machines have been extensively customized with four or five lasers and special PMTs to accommodate user protocols. Individual researchers could not support this type of high-quality cytometry in their own labs due to the high cost of the instrumentation and maintenance.